Enterobacter hormaechei KIMS8 and Enterobacter cloacae KIMS10 isolated from Kapuas River, Kalimantan, Indonesia as indigenous multi-resistant bacteria to copper and dyes




Abstract. Irawati W, Timothy M, Soentoro SE, Pinontoan R, Yuwono T, Lindarto V. 2022. Enterobacter hormaechei KIMS8 and Enterobacter cloacae KIMS10 isolated from Kapuas River, Kalimantan, Indonesia as indigenous multi-resistant bacteria to copper and dyes. Biodiversitas 23: 6661-6668. Study on the characterization of multiple copper and dyes resistance in bacteria has so far been quite limited. It is, therefore, of interest to isolate and characterize such bacteria. The study was aimed to isolate bacteria from the Kapuas River in Indonesia and measuring copper resistance as well as dye-resistance and decolorization abilities. Copper resistance was determined by measuring the minimum inhibitory concentration (MIC) of copper, while dye-resistance was measured by observing changes in colony color and clear zone formation and decolorization ability was determined by spectrophotometry. It was found that 2 out of the 15 isolated strains showed the highest copper resistance with the MIC of 7 mM. The two bacterial strains KIMS8 and KIMS10, were grown in solid media supplemented with 300 ppm of methylene blue or reactive black dye. Bacterial decolorization assays showed that KIMS8 was able to decolorize up to 90% methylene dye and up to 11.3% of reactive black dye whereas KIMS10 was able to decolorize 94.9% of methylene blue dye and 12.1% of reactive black dye. Molecular characterization by 16S rRNA gene sequencing of these two strains showed that KIMS8 and KIMS10 were identified as Enterobacter hormaechei and E. cloacae, respectively.


Abe FR, Soares AMVM, Oliveira DP, Gravato C. 2018. Toxicity of dyes to zebrafish at the biochemical level: Cellular energy allocation and neurotoxicity. Environ Pollut 235: 255–262.
An S-Y, Min S-K, Cha I-H, Choi Y-L, Cho Y-S, Kim C-H, Lee Y-C. 2002. Decolorization of triphenylmethane and azo dyes by Citrobacter sp. Biotechnol Lett 24 (12): 1037-1040.
Argüello JM, Raimunda D, Padilla-Benavides T. 2013. Mechanisms of copper homeostasis in bacteria. Front Cell Infect Microbiol 3: 73.
Azimi A, Azari A, Rezakazemi M, Ansarpour M. 2017. Removal of heavy metals from industrial wastewaters: A Review. ChemBioEng Rev 4 (1): 37–59.
The Central Bureau of Statistics of Kapuas Regency. 2018. Jumlah Perusahaan dan Tenaga Kerja menurut Jenis Industri di Kabupaten Kapuas. https://kapuaskab.bps.go.id/statictable/2015/05/25/47/jumlah-perusahaan-dan-tenaga-kerja-menurut-jenis-industri-di-kabupaten-kapuas-2013.html
Davin-Regli A, Pagès JM. 2015. Enterobacter aerogenes and Enterobacter cloacae; versatile bacterial pathogens confronting antibiotic treatment. Front Microb 6: 392.
Dupont CL, Grass G, Rensing C. 2011. Copper toxicity and the origin of bacterial resistance—new insights and applications. Metallomics 3 (11): 1109.
El Bouraie M, El Din WS. 2016. Biodegradation of Reactive Black 5 by aeromonas hydrophila strain isolated from dye-contaminated textile wastewater. Sustain Environ Res 26 (5): 209–216.
Espírito Santo C, Morais PV, Grass G. 2010. Isolation and characterization of bacteria resistant to metallic copper surfaces. Appl Environ Microbiol 76 (5): 1341–1348.
Ferreira-Leitao V, Decarvalho M, Bon E. 2007. Lignin peroxidase efficiency for methylene blue decolouration: Comparison to reported methods. Dyes Pigm 74 (1): 230–236.
Fernández-Pérez A, Marbán G. 2020. Visible light spectroscopic analysis of methylene blue in water; what comes after dimer? ACS Omega 5 (46): 29801–29815.
Fowler L, Engqvist H, Öhman-Mägi C. 2019. Effect of copper ion concentration on bacteria and cells. Materials 12 (22): 3798.
Gaetke LM, Chow-Johnson HS, Chow CK. 2014. Copper: Toxicological relevance and mechanisms. Arch Toxicol 88 (11): 1929–1938.
Giampietro R, Spinelli F, Contino M, Colabufo NA. 2018. The pivotal role of copper in neurodegeneration: A new strategy for the therapy of Neurodegenerative Disorders. Mol Pharm 15 (3): 808–820.
Grey B, Steck TR. 2001. Concentrations of copper thought to be toxic to escherichia coli can induce the viable but nonculturable condition. Appl Environ Microbiol 67 (11): 5325–5327.
Guo DJ, Singh RK, Singh P, Li DP, Sharma A, Xing YX, Song XP, Yang LT, Li YR. 2020. Complete genome sequence of Enterobacter roggenkampii ED5, a nitrogen fixing plant growth promoting endophytic bacterium with biocontrol and stress tolerance properties, isolated from sugarcane root. Front Microbiol 11: 580081.
Gou J, Liu N, Guo L, Xu H, Lv T, Yu X, Chen Y, Guo X, Rao Y, Zheng B. 2020. Carbapenem-Resistant Enterobacter hormaechei ST1103 with IMP-26 Carbapenemase and ESBL Gene blaSHV-178. Infect Drug Resist 13: 597–605.
Irawati W, Pinontoan R, Mouretta B, Yuwono T. 2022. The potential of copper-resistant bacteria Acinetobacter sp. strain CN5 in decolorizing dyes. Biodiversitas 23 (2): 680-686.
Irawati W, Tahya CY. 2021. Copper removal by Enterobacter cloacae strain IRSUK1, Enterobacter cloacae strain IRSUK4A, and Serratia nematodiphila strain IRSUK13 isolated from Sukolilo River-Indonesia. IOP Mat Sci Eng 1053 (1): 012038.
Irawati W, Djojo ES, Kusumawati L, Yuwono T, Pinontoan R. 2021. Optimizing bioremediation: Elucidating copper accumulation mechanisms of Acinetobacter sp. IRC2 isolated from an industrial waste treatment center. Front Microbiol 12: 713812.
Irawati W, Lindarto V, Pinontoan R, Yuwono T, Mangunsong F, Silalahi D. 2022. Burkholderia cepacia strain IRV1 multi-resistant to copper and dyes isolated from laboratory wastewater effluent. Biodiversitas 23 (5): 2614-2620.
Irawati W, Pinontoan R, Yuwono T. 2020. Indigenous copper resistant bacteria isolated from activated sludge of a water treatment plant in Surabaya, Indonesia. Biodiversitas 21 (11): 5077-5084.
Irawati W. 2019. Isolasi Dan Karakterisasi bakteri Resisten Tembaga dari Pantai Timur Surabaya. BIOLINK 6 (2): 95–105.
Irawati W, Parhusip AJN, Christian S, Yuwono T. 2017. The potential capability of bacteria and yeast strains isolated from Rungkut industrial sewage in Indonesia as bioaccumulators and biosorbents of copper. Biodiversitas 18 (3): 971–977.
Karim MdE, Dhar K, Hossain MdT. 2018. Decolorization of Textile Dyes by Bacterial Monoculture and Consortium Screened from Textile Dyeing Effluent. J Gen Eng Biotechnol 16 (2): 375-380.
Khandare RV, Govindwar SP. 2016. Microbial Degradation Mechanism of Textile Dye and Its Metabolic Pathway for Environmental Safety. In: Chandra, R. (1st Ed) Environmental Waste Management. CRC Press, Florida.
Khan S, Malik A. 2017. Toxicity evaluation of textile effluents and role of native soil bacterium in biodegradation of a textile dye. Environ Sci Pollut Res 25 (5): 4446–4458.
Li H-hong, Wang Y-tao, Wang Y, Wang H-xia, Sun K-kai, Lu Z-Mei. 2019. Bacterial degradation of anthraquinone dyes. J Zhejiang Univ Sci B 20 (6): 528-540.
Lellis B, Fávaro-Polonio CZ, Pamphile JA, Polonio JC. 2019. Effects of textile dyes on health and the environment and bioremediation potential of living organisms. Biotech Res Innov 3 (2): 275–290.
Lukas MC, Flitner M, Radjawali I. 2012. The conflict-laden multi-functionality of the Kapuas River in Kalimantan, Indonesia. ASEAS 5: 359–368.
Miller ME, Hamann M, Kroon FJ. 2020. Bioaccumulation and biomagnification of microplastics in marine organisms: A review and meta-analysis of current data. PLOS ONE 15 (10): e0240792.
Misal SA, Gawai KR. 2018. Azoreductase: A key player of xenobiotic metabolism. Biores Bioprocess 5 (1).
Moutaouakkil A, Zeroual Y, Zohra Dzayri F, Talbi M, Lee K, Blaghen M. 2003. Purification and partial characterization of azoreductase from Enterobacter agglomerans. Arch Biochem Biophys 413 (1): 139–146.
Nurlaila I, Irawati W, Purwandari K, Pardamean B. 2021. K-means clustering model to discriminate copper-resistant bacteria as bioremediation agents. Procedia Comp Sci 179: 804–812.
Pertile E, Vaclavik V, Dvorsky T, Heviankova S. 2020. The removal of residual concentration of hazardous metals in wastewater from a neutralization station using Biosorbent—a case study company Gutra, Czech Republic. Int J Environ Res Pub Health 17 (19): 7225.
Royer A, Sharman T. 2022. Copper Toxicity. In: StatPearls [Internet]. StatPearls Publishing, Florida.
Shamim S. 2018. Biosorption of heavy metals. In: Derco, J., Vrana, B., Biosorption. IntechOpen, London.
Tkaczyk A, Mitrowska K, Posyniak A. 2020. Synthetic organic dyes as contaminants of the aquatic environment and their implications for ecosystems: A review. Sci Tot Environ 717: 137222.
Tyt?a M, Widziewicz K, Zielewicz E. 2015. Heavy metals and its chemical speciation in sewage sludge at different stages of processing. Environ Technol 37 (7): 899–908.
Varma VG, Misra AK. 2016. Copper contaminated wastewater – an evaluation of bioremedial options. Indoor Built Environ 27 (1): 84–95.
Wielewski LP, Zuccolotto T, Soares M, Prola LDT. 2020. Degradation of the Textile Dye Reactive Black 5 by Basidiomycetes. Rev Ambient Agua 15 (1).
Zucca P, Cocco G, Sollai F, Sanjust E. 2016. Fungal laccases as tools for biodegradation of industrial dyes. Biocatalysis 1 (1).

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